JP6771161B2 - Nucleic acid extractor - Google Patents

Description

The present invention relates to a nucleic acid extraction device for extracting nucleic acid (deoxyribonucleic acid, ribonucleic acid) from a sample, a nucleic acid extraction unit used in the nucleic acid extraction device, and a nucleic acid extraction method for extracting nucleic acid from a sample.

Nucleic acid amplification methods such as polymerase chain reaction (PCR) are used in the examination of microorganisms such as bacteria contained in beverages or foods. The nucleic acid amplification method has an advantage that the test process can be significantly speeded up and simplified as compared with the culture method.

In order to carry out a test using the nucleic acid amplification method, a pretreatment for extracting nucleic acid from a sample is required before performing nucleic acid amplification. Conventionally, as such a pretreatment, a method of capturing a test object such as bacteria contained in the sample by filtering the sample (sample stock solution) with a filter and extracting nucleic acid from the test object with a nucleic acid extraction reagent has been used. It is known (for example, Patent Document 1).

Japanese Unexamined Patent Publication No. 4-36197

In the conventional nucleic acid extraction method (pretreatment) disclosed in Patent Document 1, after filtering and concentrating the test object through a syringe containing a sample, this filter is removed and another nucleic acid containing a nucleic acid extraction reagent is used. It is reattached to elute the test object and extract the nucleic acid.

However, in the conventional nucleic acid extraction method, the work is complicated, and there is a risk that impurities such as bacteria different from the test object may be mixed in the nucleic acid extract containing the extracted nucleic acid (contamination) during the work. is there.

The present invention has been made to solve such a problem, and provides a nucleic acid extraction device, a nucleic acid extraction unit, and the like, which are easy to work with and can significantly reduce the risk of impurities being mixed in the nucleic acid extract. The purpose is.

In order to achieve the above object, one aspect of the nucleic acid extraction device according to the present invention includes a nucleic acid extraction unit for extracting nucleic acid from a sample, and the nucleic acid extraction unit injects a container and a sample into the container. A first injection port for the purpose, at least a part thereof is arranged in the container, and a capture unit for capturing a nucleic acid extraction target contained in a sample injected from the first injection port, and a capture unit captured by the capture unit. A second injection port for injecting a nucleic acid extraction reagent for extracting nucleic acid from the nucleic acid extraction target into the container, and a nucleic acid containing the nucleic acid extracted from the nucleic acid extraction target by the nucleic acid extraction reagent. The container has a first discharge port for discharging the extract from the container and a second discharge port for discharging the sample from the container, and the container has an internal space of the container with the catching portion as a boundary. The first container portion and the second container portion are provided, and the first injection port and the second injection port are provided in the first container portion, and the first discharge port and the second injection port are provided. The discharge port is provided in the second container portion.

Further, one aspect of the nucleic acid extraction unit according to the present invention is a nucleic acid extraction unit used in a nucleic acid extraction device for extracting nucleic acid from a sample, and is a container and a first for injecting a sample into the container. The inlet, the capture unit that is placed in the container and captures the nucleic acid extraction target contained in the sample injected from the first injection port, and the capture unit that captures the nucleic acid from the nucleic acid extraction target captured by the capture unit. To discharge a second injection port for injecting a nucleic acid extraction reagent for extraction into the container and a nucleic acid extract containing the nucleic acid extracted from the nucleic acid extraction target by the nucleic acid extraction reagent from the container. The container is provided with a first discharge port and a second discharge port for discharging the sample from the container, and the container has a first container portion for dividing the internal space of the container with the capture portion as a boundary. It has a second container portion, the first injection port and the second injection port are provided in the first container portion, and the first discharge port and the second discharge port are the second container. It is provided in the section.

Further, one aspect of the nucleic acid extraction method according to the present invention is a capture step of capturing a nucleic acid extraction target contained in the sample by the capture unit by passing the sample through the capture unit, and a nucleic acid extraction reagent in the capture unit. Includes a nucleic acid extraction step of extracting nucleic acid from the nucleic acid extraction target captured by the capture unit by introducing the above, and a recovery step of collecting the nucleic acid extract containing the extracted nucleic acid through the capture unit. ..

According to the present invention, workability is simple, and the risk of impurities being mixed in the nucleic acid extract can be significantly reduced.

FIG. 1 is a perspective view schematically showing a nucleic acid extraction device according to an embodiment. FIG. 2 is an enlarged perspective view of a main part showing the peripheral structure of the nucleic acid extraction unit in the nucleic acid extraction device according to the embodiment. FIG. 3 is a perspective view of the nucleic acid extraction unit according to the embodiment. FIG. 4 is a cross-sectional view of the nucleic acid extraction unit according to the embodiment. FIG. 5 is a flowchart of the nucleic acid extraction method according to the embodiment. FIG. 6A is a diagram showing a state when a sample is injected into a container in the nucleic acid extraction method according to the embodiment. FIG. 6B is a diagram showing a nucleic acid extraction target captured by the capture unit in the nucleic acid extraction method according to the embodiment. FIG. 6C is a diagram showing a state when a nucleic acid extraction reagent is injected into a container in the nucleic acid extraction method according to the embodiment. FIG. 6D is a diagram showing a state when the nucleic acid extraction reagent is held in the capture unit in the nucleic acid extraction method according to the embodiment. FIG. 6E is a diagram showing a state when the nucleic acid extract is moved from the first container portion to the second container portion in the nucleic acid extraction method according to the embodiment. FIG. 6F is a diagram showing a state of the nucleic acid extract liquid moved to the second container portion in the nucleic acid extraction method according to the embodiment. FIG. 6G is a diagram showing a state when the nucleic acid extract is discharged from the container in the nucleic acid extraction method according to the embodiment. FIG. 7 is a cross-sectional view of the nucleic acid extraction unit according to the first modification. FIG. 8 is a cross-sectional view of the nucleic acid extraction unit according to the second modification. FIG. 9 is a cross-sectional view of the nucleic acid extraction unit according to the third modification. FIG. 10 is a cross-sectional view of the nucleic acid extraction unit according to the modified example 4. FIG. 11 is a cross-sectional view of the nucleic acid extraction unit according to the modified example 5.

Hereinafter, embodiments of the present invention will be described with reference to the drawings. It should be noted that all of the embodiments described below show a preferred specific example of the present invention. Therefore, the numerical values, shapes, materials, components, the arrangement positions and connection forms of the components, the steps (processes), the order of the steps, and the like shown in the following embodiments are examples and limit the present invention. It is not the purpose of doing it. Therefore, among the components in the following embodiments, the components not described in the independent claims indicating the highest level concept of the present invention will be described as arbitrary components.

It should be noted that each figure is a schematic view and is not necessarily exactly illustrated. Further, in each figure, the same reference numerals are given to substantially the same configurations, and duplicate description will be omitted or simplified.

(Embodiment)
[Nucleic acid extractor]
The configurations of the nucleic acid extraction device 100 and the nucleic acid extraction unit 1 according to the embodiment will be described with reference to FIGS. 1 to 4. FIG. 1 is a perspective view schematically showing a nucleic acid extraction device 100 according to an embodiment. FIG. 2 is an enlarged perspective view of a main part showing the peripheral structure of the nucleic acid extraction unit 1 in the nucleic acid extraction device 100, and shows the nucleic acid extraction unit 1 in a state of being installed in the nucleic acid extraction device 100. FIG. 3 is a perspective view of the nucleic acid extraction unit 1 according to the embodiment, and FIG. 4 is a cross-sectional view of the nucleic acid extraction unit 1.

As shown in FIG. 1, the nucleic acid extraction device 100 includes a nucleic acid extraction unit 1 for extracting nucleic acid from an object to be measured contained in a sample, a vacuum pump 2, and a vibration device 3.

The object to be measured contained in the sample (sample stock solution) is an object for extracting nucleic acid (nucleic acid extraction object), and is, for example, a microorganism such as a bacterium, a virus, or a tissue cell. The sample stock solution containing microorganisms can be collected from, for example, a beverage.

Microorganisms such as bacteria, viruses, and histiocytes are test objects that are tested by nucleic acid amplification such as PCR. That is, the nucleic acid extraction device 100 is a device used for pretreatment before performing nucleic acid amplification, and the nucleic acid extracted by the nucleic acid extraction device 100 is used for a desired test. In the nucleic acid extraction device 100 of the present embodiment, the nucleic acid is recovered as a liquid (nucleic acid extract) containing the extracted nucleic acid.

The nucleic acid extraction unit 1 is a processing unit for extracting nucleic acid from a sample, and is installed in the reaction box 4 of the nucleic acid extraction device 100 as shown in FIG. In this embodiment, the nucleic acid extraction unit 1 is a replaceable cartridge and can be removed from the reaction box 4. The nucleic acid extraction unit 1 is replaced, for example, in each nucleic acid extraction process. Note that FIG. 1 shows a state in which the entire lid of the reaction box 4 is opened.

As shown in FIGS. 3 and 4, the nucleic acid extraction unit 1 includes a container 10, a first injection port 21, a second injection port 22, a first discharge port 31, a second discharge port 32, and a capture unit. 40 and.

The container 10 is a processing container for extracting nucleic acid from the sample by the sample to be injected and the nucleic acid extraction reagent. The nucleic acid extracted from the sample becomes a nucleic acid extract and is discharged from the container 10.

The material of the container 10 is not particularly limited, but the container 10 has high heat resistance such as polypropylene (PP) or polycarbonate (PC) so that the heat treatment can be performed by the nucleic acid extraction treatment. It may be composed of a resin material, a metal material such as aluminum or stainless steel, or an inorganic material such as glass or ceramic. Further, when a resin material is used as the container 10, there is an advantage that the weight can be reduced at low cost. Further, when a metal material or a highly thermally conductive resin is used as the container 10, the thermal conductivity of the container 10 can be improved. Therefore, when heat treatment is performed, it is preferable that at least a part of the container 10 is made of a metal material or a highly thermally conductive resin. The container 10 may be formed by combining the above materials.

Further, the inner surface of the container 10 is preferably non-hydrophilic. For example, the inner surface of the container 10 may be hydrophobic. In this case, the container 10 may be formed by using a hydrophobic material, the inner surface of the container 10 may be made hydrophobic by surface treatment, or a hydrophobic film is coated on the inner surface of the container 10. May be good.

The container 10 is composed of a first container portion 11 and a second container portion 12. The first container portion 11 and the second container portion 12 divide the internal space of the container 10 with the catching portion 40 as a boundary. In the present embodiment, the container 10 is divided into two in the vertical direction with the main body 41 of the catching portion 40 as a boundary, the first container portion 11 is the upper component, and the second container portion 12 is the lower component. It has become. The first container portion 11 is further divided into two, but the present invention is not limited to this.

The first injection port 21, the second injection port 22, the first discharge port 31, the second discharge port 32, and the capture unit 40 are provided in the container 10. Specifically, the first injection port 21, the second injection port 22, the first discharge port 31, and the second discharge port 32 are provided on the partition wall of the container 10, and the capture unit 40 has the main body 41 as the container. It is fixed to the container 10 so as to be located inside the 10.

The first injection port 21 is a sample injection port for injecting a sample into the container 10. In the present embodiment, the first injection port 21 is provided on the upper wall of the first container portion 11. The sample (sample stock solution) is injected into the first container portion 11 from the sample charging cup 200 installed in the nucleic acid extraction device 100 via the first injection port 21.

The first injection port 21 is a pipe that communicates the outside of the container 10 with the internal space of the first container portion 11. Specifically, one end of the first injection port 21 is connected to the first container portion 11, and the other end of the first injection port 21 is connected to the sample charging cup 200. In the present embodiment, the first injection port 21 is integrally formed with the first container portion 11. The first injection port 21 may be a through hole provided in the upper wall of the container 10 (first container portion 11).

The second injection port 22 is a nucleic acid extraction reagent injection port for injecting the nucleic acid extraction reagent into the container 10. In the present embodiment, the second injection port 22 is provided on the upper wall of the first container portion 11 like the first injection port 21, but is provided at a position different from that of the first injection port 21.

The nucleic acid extraction reagent injected from the second injection port 22 is a liquid reagent for extracting nucleic acid from the nucleic acid extraction target captured by the capture unit 40, and for example, has an action of extracting nucleic acid from the cell membrane of the nucleic acid extraction target. Have. The nucleic acid extraction reagent is injected into the first container portion 11 from the nucleic acid extraction reagent container 300 installed in the nucleic acid extraction device 100 via the second injection port 22.

The second injection port 22 is a through hole provided in the first container portion 11. The second injection port 22, which is a through hole, is provided with a rubber stopper 22a that closes the second injection port 22 (through hole). The rubber stopper 22a is made of a rubber material through which a needle for injecting the nucleic acid extraction reagent into the first container portion 11 can penetrate. When the nucleic acid extraction reagent in the nucleic acid extraction reagent container 300 is injected into the first container portion 11, the needle provided in the nucleic acid extraction reagent container 300 is passed through the rubber stopper 22a. As a result, the liquid reagent can be injected from the nucleic acid extraction reagent container 300 into the first container portion 11 via the second injection port 22. The rubber material of the rubber stopper 22a is not particularly limited, but for example, silicone rubber, fluororubber, or the like is used.

Since the liquid of the sample and the nucleic acid extraction reagent is injected into the first container portion 11 in this way, the angle of the corner portion on the wall surface of the internal space of the first container portion 11 is preferably 90 degrees or more. That is, it is preferable that the angle formed by any two wall surfaces in the first container portion 11 is 90 degrees or more. Thereby, when the sample and the nucleic acid extraction reagent are injected into the first container portion 11, it is possible to prevent the sample and the nucleic acid extraction reagent from staying on the wall surface of the internal space of the first container portion 11.

The first discharge port 31 is a nucleic acid extract discharge port for discharging the nucleic acid extract containing the nucleic acid extracted from the nucleic acid extraction target by the nucleic acid extraction reagent from the container 10. In the present embodiment, the first discharge port 31 is provided on the lower wall of the second container portion 12.

The first discharge port 31 is a pipe that communicates the outside of the container 10 with the internal space of the second container portion 12. Specifically, one end of the first discharge port 31 was connected to the second container portion 12, and the other end of the first discharge port 31 was installed in the nucleic acid extraction device 100 via a pipe. It is connected to a recovery container 400 (nucleic acid extract recovery container). In the present embodiment, the first discharge port 31 is integrally formed with the second container portion 12.

The nucleic acid extract discharged from the first discharge port 31 is collected in the collection container 400 through a pipe connected to the first discharge port 31. For example, the nucleic acid extract is recovered by being pumped from the container 10 to the recovery container 400 by the liquid feed pump 5.

The second discharge port 32 is a sample discharge port for discharging the sample from the container 10. The waste liquid of the sample injected into the container 10 is discharged from the second discharge port 32. That is, the sample (filter solution) that has been injected from the first container unit 11 and passed through the capture unit 40 is discharged from the second discharge port 32. In the present embodiment, the second discharge port 32 is provided on the side wall of the second container portion 12. That is, the second discharge port 32 is provided at a position closer to the catching portion 40 than the first discharge port 31.

Further, the opening diameter (inner diameter) of the second discharge port 32 is preferably twice or more the opening diameter (inner diameter) of the first discharge port 31. Generally, the waste liquid of the sample has a large volume (for example, about 100 ml), and the nucleic acid extract recovered for nucleic acid amplification has a small volume (for example, about 200 μl). Therefore, as in the present embodiment, the discharge route between the waste liquid of the sample and the nucleic acid extract is divided into the first discharge port 31 and the second discharge port 32, and the opening diameter of the second discharge port 32 is set to the first. By setting the opening diameter of the discharge port 31 to twice or more, it is possible to reduce losses such as the nucleic acid extract adhering to the wall surface of the container 10. That is, by reducing the opening diameter of the first discharge port 31, the nucleic acid extract can be discharged with less loss. For example, the opening diameter of the second discharge port 32 is 2 mm, and the opening diameter of the first discharge port 31 is 0.5 mm. The opening diameter of the second discharge port 32 does not have to be twice or more the opening diameter of the first discharge port 31, and the opening diameter of the second discharge port 32 is the same as the opening diameter of the first discharge port 31. It may be smaller than the opening diameter of the first discharge port 31.

The second discharge port 32 is a pipe that communicates the outside of the container 10 with the internal space of the second container portion 12. Specifically, one end of the second discharge port 32 is connected to the second container part 12, and the other end of the second discharge port 32 is installed in the nucleic acid extraction device 100 via a pipe. It is connected to the collection container 500 (sample waste liquid collection container). In the present embodiment, the second discharge port 32 is integrally formed with the second container portion 12.

The sample discharged from the second discharge port 32 is collected in the collection container 500 through the pipe connected to the second discharge port 32. For example, the waste liquid of the sample is collected by being sent from the container 10 to the collection container 500 by the vacuum pump 2.

The capture unit 40 is a capture unit for capturing the nucleic acid extraction target contained in the sample injected from the first injection port 21, and at least a part thereof is arranged in the container 10. In the present embodiment, the capture unit 40 captures the nucleic acid extraction target contained in the sample by filtration.

The capture unit 40 has a main body portion 41 for capturing and holding the nucleic acid extraction target, and a support portion 42 for supporting the main body portion 41. The catching portion 40 is fixed to the container 10 so that the main body portion 41 is located inside the container 10.

The main body 41 is placed on the plate-shaped support 42. A plurality of through holes (eyes) are formed in the central portion of the support portion 42 corresponding to the main body portion 41. That is, the support portion 42 is a perforated plate. Further, the peripheral portion of the support portion 42 is sandwiched between the first container portion 11 and the second container portion 12. As a result, the support portion 42 is fixed to the container 10. The support portion 42, the first container portion 11, and the second container portion 12 are fixed by, for example, four screws.

In the present embodiment, the main body 41 is a filter for filtering and capturing a nucleic acid extraction target (microorganism) from a sample. Specifically, the main body 41 is a filter having a plurality of fine pores (eyes) smaller than the size of the nucleic acid extraction target. By this. The main body 41 (filter) can reliably capture the nucleic acid extraction target. The hole diameter of the main body 41 is, for example, 0.45 μm. The nucleic acid extraction target captured by the main body 41 is held on the main body 41. Further, in the present embodiment, since the nucleic acid extracted from the nucleic acid extraction target is passed through the main body 41, the pore size of the main body 41 may be made larger than the size of the nucleic acid.

As the main body 41, a membrane filter made of a material such as cellulose acetate, polyvinylidene fluoride (PVDF), polyethersulfone (PES), or cellulose acetate can be used. Further, as the main body 41, a capture filter having a function of adsorbing a nucleic acid extraction target contained in a sample may be used. By using such a capture filter, the rate of capturing the nucleic acid extraction target from the sample can be improved. In the present embodiment, the main body 41 (filter) is a flat filter, but the present invention is not limited to this, and a filter having another shape such as a cylindrical shape may be used.

At least a part of the support portion 42 may be made of a metal material such as aluminum or a high thermal conductive resin. In this case, the support portion 42 may be entirely made of a metal material or a high thermal conductive resin. When a metal material is used for a part of the support portion 42, the metal material may be embedded in the resin. Further, the surface of the support portion 42 is preferably hydrophilic.

The vacuum pump 2 is an example of a pressure adjusting unit for adjusting the pressure in the container 10. In the present embodiment, the piping of the vacuum pump 2 is connected to the second container portion 12. Therefore, when the liquid is accumulated on the trapping portion 40 and the internal space of the second container portion 12 is sealed, the vacuum pump 2 adjusts the pressure in the second container portion 12. The pressure in the container 10 or in the second container portion 12 can be adjusted by the leak valve 6 provided in the piping of the vacuum pump 2. That is, the degree of decompression by the vacuum pump 2 can be adjusted by the leak valve 6.

Further, in the present embodiment, the piping of the vacuum pump 2 is connected to the second discharge port 32. That is, the decompression port (decompression suction port) of the vacuum pump 2 also serves as the second discharge port 32 provided in the second container portion 12.

The vibrating device 3 has a function for vibrating the container 10. By vibrating the container 10 with the vibrating device 3, the liquid in the container 10 can be agitated. For example, the nucleic acid extraction reaction can be effectively carried out by stirring the nucleic acid extraction reagent containing the nucleic acid extraction target. The vibrating device 3 is provided so as to come into contact with the reaction box 4.

[Nucleic acid extraction method]
Next, the nucleic acid extraction method using the nucleic acid extraction device 100 will be described with reference to FIGS. 5 and 6A to 6G with reference to FIG. FIG. 5 is a flowchart of the nucleic acid extraction method according to the embodiment. 6A to 6G are schematic cross-sectional views for explaining the nucleic acid extraction method according to the embodiment. In FIGS. 6A to 6G, the first injection port 21 and the second injection port 22 (rubber stopper 22a) are shown at correct positions in order to make it easier to understand the flow of the liquids of the sample 210 and the nucleic acid extraction reagent. Not.

The nucleic acid extraction method in the present embodiment is a method of extracting nucleic acid from a nucleic acid extraction target contained in a sample by using the nucleic acid extraction unit 1, and as shown in FIG. 5, at least the capture step S1 and the nucleic acid extraction. A step S2 and a recovery step S3 are included.

The capture step S1 is a step in which the capture unit 40 captures the nucleic acid extraction target contained in the sample by passing the sample through the capture unit 40.

The nucleic acid extraction step S2 is a step of extracting nucleic acid from the nucleic acid extraction target captured by the capture unit 40 by introducing a nucleic acid extraction reagent into the capture unit 40 after the capture step S1.

The recovery step S3 is a step of recovering the nucleic acid extract containing the nucleic acid extracted in the nucleic acid extraction step S2 by passing it through the capture unit 40 after the nucleic acid extraction step S2.

Hereinafter, a specific nucleic acid extraction method will be described with reference to FIGS. 6A to 6G.

[Capture process]
In the capture step S1, first, as shown in FIG. 6A, the sample 210 is injected into the nucleic acid extraction unit 1. Specifically, a sample charging cup 200 containing a sample 210 (sample stock solution) containing a nucleic acid extraction target 220 is installed at a predetermined position of the nucleic acid extraction device 100 (see FIG. 1), and a container is placed from the sample charging cup 200. 10 is charged into the sample 210. When the sample 210 is put into the container 10, the valve 5a provided in the pipe connected to the first discharge port 31 is closed. The valve 5a is provided in, for example, the liquid feed pump 5 in FIG. 1, but is not limited thereto.

The sample 210 can be prepared, for example, by suspending a sample containing a microorganism in sterile diluted water or the like. The amount of the solution of the sample 210 is, for example, 10 ml to 500 ml, but is not limited to this. If necessary, the sample 210 may be subjected to pretreatment such as removal of solid components.

The sample 210 charged from the sample charging cup 200 is injected into the first container section 11 through the first injection port 21, passes through the capturing section 40, and flows into the second container section 12.

At this time, the sample 210 is concentrated by the capture unit 40. That is, the sample 210 can be concentrated by passing the sample 210 through the capture unit 40.

Specifically, when the sample 210 passes through the capture unit 40, the nucleic acid extraction target 220 contained in the sample 210 is captured by the capture unit 40. In the present embodiment, the nucleic acid extraction target 220 contained in the sample 210 is captured by the main body 41 when passing through the main body 41 (filter) of the capture unit 40 and stays on the main body 41.

As shown in FIG. 6A, the sample 210 (filter solution) that has passed through the capture unit 40 is discharged from the second container unit 12 as a waste liquid through the second discharge port 32, and is discharged from the nucleic acid extraction device 100 (see FIG. 1). It is collected in a collection container 500 installed at a predetermined position.

At this time, in the present embodiment, the nucleic acid extraction target 220 contained in the sample 210 is captured by the capture unit 40 by suction filtration. Specifically, the vacuum pump 2 exhausts (sucks) the air in the container 10 from the first discharge port 31 to reduce the pressure in the second container portion 12 and reduce the pressure in the first container portion 11 to the second. The pressure inside the second container portion 12 is adjusted in the sample discharge mode in which the pressure is higher than the pressure inside the container portion 12. By adjusting the pressure in the container 10 in this way, the sample 210 injected into the container 10 is filtered by the capture unit 40 while being sucked. As a result, the sample 210 injected into the first container section 11 can be quickly passed through the capture section 40 and moved to the second container section 12, and the waste liquid of the sample 210 can be quickly moved from the second container section 12. Can be discharged. Therefore, the sample 210 can be efficiently concentrated, and the waste liquid of the sample 210 can be quickly discharged from the container 10.

When concentrating the sample 210, the sample 210 may be further pressurized from above. As a result, the sample 210 can be concentrated more efficiently. For example, the space area above the sample 210 in the sample charging cup 200 may be pressurized.

When the sample 210 is injected into the container 10 and discharged in this way, as shown in FIG. 6B, the nucleic acid extraction target 220 captured by the main body 41 accumulates on the main body 41 of the capture unit 40. It will be.

In this way, in the capture step S1, the capture unit 40 is allowed to capture the nucleic acid extraction target 220 contained in the sample 210 by passing the sample 210 through the capture unit 40.

After passing the sample 210 through the capture unit 40, if necessary, sterilized diluted water is further introduced into the first container unit 11 from the first injection port 21 or the like and passed through the capture unit 40 to pass the nucleic acid. The extraction target 220 can be washed.

[Nucleic acid extraction process]
In the nucleic acid extraction step S2, the nucleic acid 230 is extracted from the nucleic acid extraction target 220 captured by the capture unit 40.

Specifically, first, as shown in FIG. 6C, the nucleic acid extraction reagent 310 is charged into the container 10 from the nucleic acid extraction reagent container 300 installed at a predetermined position of the nucleic acid extraction device 100 (see FIG. 1). As the nucleic acid extraction reagent 310, for example, a simple DNA extraction kit (Kaneka Corporation) or Celeas (Biocosm Co., Ltd.) can be used. The amount of the nucleic acid extraction reagent 310 to be injected is, for example, 50 μl to 200 μl. When the nucleic acid extraction reagent 310 is charged into the container 10, the valve 5a of the pipe connected to the first discharge port 31 remains closed.

The nucleic acid extraction reagent 310 charged from the nucleic acid extraction reagent container 300 is injected into the first container section 11 via the second injection port 22. Specifically, the needle of the nucleic acid extraction reagent container 300 is passed through the rubber stopper 22a provided in the second injection port 22 (through hole), and the nucleic acid extraction reagent is penetrated from the nucleic acid extraction reagent container 300 into the first container portion 11. Inject 310. Before injecting the nucleic acid extraction reagent 310 into the container 10, an inert treatment reagent for killed bacteria or the like may be injected into the container 10.

When the nucleic acid extraction reagent 310 is injected into the container 10 and allowed to stand, the nucleic acid extraction target 220 captured by the capture unit 40 reacts with the nucleic acid extraction reagent 310. Thereby, the nucleic acid 230 can be extracted from the nucleic acid extraction target 220. That is, a nucleic acid extraction reaction is performed in which the nucleic acid 230 is extracted from the nucleic acid extraction target 220. That is, the cell membrane of the nucleic acid extraction target 220 and the nucleic acid 230 are separated. Specifically, as shown in FIG. 6D, the nucleic acid 230 is eluted from the nucleic acid extraction target 220 into the nucleic acid extraction reagent 310 to generate a nucleic acid extract 320 containing the nucleic acid 230.

The nucleic acid extraction reagent 310 may be injected into the container 10 to mix the nucleic acid extraction target 220 and the nucleic acid extraction reagent 310, and then appropriately heated. As a result, the reaction between the nucleic acid extraction reagent 310 and the nucleic acid extraction target 220 can be promoted to efficiently elute the nucleic acid 230, so that the nucleic acid 230 can be extracted efficiently. In this case, for example, the nucleic acid extraction reagent 310 can be heated by heating the container 10 with the heating / cooling unit 7 shown in FIG. The heating / cooling unit 7 is provided in, for example, the reaction box 4 of the nucleic acid extraction device 100, and is connected to, for example, the container 10 and the support portion 42 of the capture portion 40. As the heating / cooling unit 7, for example, a Peltier element or the like can be used.

Further, after mixing the nucleic acid extraction target 220 and the nucleic acid extraction reagent 310, the container 10 may be vibrated by the vibrating device 3 to stir the nucleic acid extraction reagent 310. As a result, the extraction of nucleic acid 230 can be performed more efficiently.

As described above, in the present embodiment, the nucleic acid extraction reaction of the nucleic acid extraction target 220 is performed by introducing the nucleic acid extraction reagent 310 into the capture unit 40, and the nucleic acid 230 is extracted from the nucleic acid extraction target 220. As a result, a nucleic acid extract 320 containing the extracted nucleic acid 230 is generated.

At this time, as shown in FIG. 6D, the nucleic acid extraction reaction of the nucleic acid extraction target 220 may be carried out by holding the nucleic acid extraction reagent 310 injected into the container 10 on the capture unit 40. By holding the nucleic acid extraction reagent 310 on the capture unit 40, the nucleic acid extraction reagent 310 stays on the capture unit 40. As a result, the nucleic acid extraction target 220 is immersed in the nucleic acid extraction reagent 310 for a certain period of time. As a result, the nucleic acid 230 can be easily eluted, so that the nucleic acid 230 can be efficiently extracted (eluted).

When the nucleic acid extraction reagent 310 is held on the capture unit 40, the vacuum pump 2 adjusts the pressure in the second container unit 12 in the nucleic acid extraction reagent holding mode in which the nucleic acid extraction reagent 310 is held on the capture unit 40. Good. Specifically, the inside of the second container portion 12 may be pressurized by the vacuum pump 2 so that the pressure inside the second container portion 12 is higher than the pressure inside the first container portion 11. As a result, the nucleic acid extraction reagent 310 injected into the container 10 can be held on the capture unit 40.

In this case, as the main body 41 of the capture unit 40, a filter can be used so that the nucleic acid extraction reagent 310 passes through the main body 41 in the normal state (when the second container 12 is not pressurized).

[Recovery process]
After the injection of the nucleic acid extraction reagent 310 is completed and the nucleic acid extraction reaction of the nucleic acid extraction target 220 on the capture unit 40 is completed, the pressurization in the second container unit 12 by the vacuum pump 2 is stopped. As shown in 6E, the nucleic acid extract 320 staying on the capture unit 40 can be moved to the second container unit 12 through the capture unit 40. For example, by opening the leak valve 6, the pressure inside the second container 12 can be adjusted to normal pressure, or the pressure inside the second container 12 can be adjusted so that the pressure is lower than the pressure reduced in the sample discharge mode. You can do it.

The method of holding the nucleic acid extraction reagent 310 on the trapping unit 40 is not limited to the method of adjusting the pressure in the second container unit 12 as described above. For example, instead of executing the nucleic acid extraction reagent holding mode, as the main body 41 of the capture unit 40, a filter such that the nucleic acid extraction reagent 310 is held on the main body 41 when the inside of the second container 12 is not depressurized. It can also be realized by using.

In this case, although not shown, after the nucleic acid extraction reaction of the nucleic acid extraction target 220 on the capture unit 40 is completed by the nucleic acid extraction reagent 310 held on the capture unit 40, the second container unit 12 is operated by the vacuum pump 2. The pressure inside the first container portion 11 may be reduced to be higher than the pressure inside the second container portion 12. As a result, the nucleic acid extract 320 that has completed the nucleic acid extraction reaction and remains on the capture unit 40 can be moved to the second container unit 12. That is, the nucleic acid extract 320 can be moved to the second container portion 12 by utilizing the pressure difference. In this way, the pressure inside the second container portion 12 is reduced by the vacuum pump 2 to make the pressure inside the first container portion 11 higher than the pressure inside the second container portion 12, so that the nucleic acid extract 320 in the capture unit 40 It is preferable to adjust the pressure in the second container portion 12 in the nucleic acid extract moving mode in which the is moved to the second container portion 12 via the trapping portion 40.

At this time, the degree of decompression by the vacuum pump 2 in the nucleic acid extract transfer mode may be lower than the degree of decompression by the vacuum pump 2 in the sample discharge mode in which the sample 210 is discharged.

The second container portion 12 may be controlled so that a centrifugal force is applied in the sample discharge mode and the nucleic acid extract movement mode. As a result, the sample 210 and the nucleic acid extract 320 can be effectively recovered.

Next, as shown in FIG. 6F, after all the nucleic acid extract 320 containing the nucleic acid 230 has moved to the second container portion 12, as shown in FIG. 6G, the pipe connected to the first discharge port 31 The nucleic acid extract 320 is recovered by opening the valve 5a.

Specifically, the nucleic acid extract 320 containing the nucleic acid 230 is discharged from the second container portion 12 via the first discharge port 31, and is collected at a predetermined position of the nucleic acid extraction device 100 (see FIG. 1). The nucleic acid extract 320 is collected in the container 400. For example, the nucleic acid extract 320 in the second container portion 12 is sent to the recovery container 400 by the liquid feed pump 5 and collected.

Then, although not shown, a nucleic acid test (nucleic acid analysis) is performed using the recovered nucleic acid extract 320. In this case, a nucleic acid test reagent is mixed with the nucleic acid extract 320 according to the method for testing the nucleic acid 230 to obtain a test solution. For example, when a nucleic acid test is performed by the PCR method, a PCR reagent is used as the nucleic acid test reagent. The PCR reagent includes, for example, a nucleic acid-staining fluorescent reagent for examining the amount of nucleic acid 230 by fluorescence emission intensity, a reaction reagent for amplifying nucleic acid, and the like. The reaction reagent is, for example, a PCR primer, a polymerase enzyme, a buffer, or the like.

The nucleic acid test by the PCR method can be performed, for example, by using a test chip (PCR chip or the like) in which a microchannel, which is a test channel, is formed. In this case, for example, nucleic acid extraction is performed by introducing a mixed solution of the nucleic acid extract 320 and the nucleic acid test reagent into the test chip, amplifying the nucleic acid 230 by flow PCR, and measuring the nucleic acid 230 amplified by the optical detection device. The object 220 (nucleic acid) can be identified.

As described above, the nucleic acid extraction method in the present embodiment can be used as a nucleic acid test method. In this case, by examining the nucleic acid 230 by the PCR method, the nucleic acid 230 can be identified with high sensitivity and high accuracy.

[Summary]
As described above, according to the nucleic acid extraction device 100 according to the present embodiment, the nucleic acid extraction unit 1 for extracting the nucleic acid 230 from the sample 210 is provided. The nucleic acid extraction unit 1 is arranged in the container 10, the first injection port 21 for injecting a sample into the container 10, and the nucleic acid extraction contained in the sample 210 injected from the first injection port 21. A capture unit 40 that captures the object 220, and a second injection port 22 for injecting a nucleic acid extraction reagent 310 for extracting nucleic acid 230 from the nucleic acid extraction target 220 captured by the capture unit 40 into the container 10. A first discharge port 31 for discharging the nucleic acid extract 320 containing the nucleic acid 230 extracted from the nucleic acid extraction target 220 by the nucleic acid extraction reagent 310 from the container 10, and a first discharge port 31 for discharging the sample 210 from the container 10. It has two discharge ports 32. The container 10 has a first container portion 11 and a second container portion 12 that divide the internal space of the container 10 with the catching portion 40 as a boundary, and the first injection port 21 and the second injection port 22 are second. 1 The container portion 11 is provided, and the first discharge port 31 and the second discharge port 32 are provided in the second container portion 12.

As a result, the sample 210 and the nucleic acid extraction reagent 310 can be injected into the upper first container portion 11, and the waste liquid sample 210 and the nucleic acid extract 320 can be injected from the lower second container portion 12 via the capture unit 40. And can be discharged. As a result, the nucleic acid extract 320 containing the nucleic acid 230 can be easily recovered after concentrating the sample 210 and extracting the nucleic acid 230 using the same container 10. Further, since the work can be performed without transferring the container during the nucleic acid extraction step, it is possible to prevent impurities from being mixed in the nucleic acid extract 320. Moreover, since the sample 210 flows from the upper first container portion 11 to the lower second container portion 12, the nucleic acid extraction target 220 can be easily captured by the capture unit 40 from the sample 210. As described above, according to the present embodiment, the workability is simple and the risk of impurities being mixed in the nucleic acid extract 320 can be significantly reduced. As a result, nucleic acid 230 can be extracted with high sensitivity and high accuracy. Therefore, the accuracy of the test using the nucleic acid extract 320 can be improved.

Further, in the present embodiment, the nucleic acid extraction device 100 further includes a vacuum pump 2 as a pressure adjusting unit for adjusting the pressure in the second container unit 12. Further, the capture unit 40 has a filter having a plurality of pores smaller than the nucleic acid extraction target 220 as the main body unit 41. The second discharge port 32 is provided at a position closer to the capture unit 40 than the first discharge port 31, and also serves as a decompression port of the vacuum pump 2.

As a result, the nucleic acid extract 320 can be efficiently recovered even though the nucleic acid extract 320 required for the test and the sample 210 to be waste liquid are discharged from the same second container portion 12.

Further, in the present embodiment, the vacuum pump 2 is injected by reducing the pressure in the second container portion 12 and making the pressure in the first container portion 11 higher than the pressure in the second container portion 12. A sample discharge mode in which the sample 210 is moved from the first container section 11 through the capture section 40 to the second container section 12, and the pressure inside the second container section 12 is reduced to reduce the pressure inside the first container section 11. In the nucleic acid extract moving mode in which the nucleic acid extract 320 in the trapping unit 40 is moved to the second container 12 via the trapping unit 40 by making the pressure higher than the pressure in the container portion 12, the pressure in the second container portion 12 Adjust the pressure. In this case, the degree of decompression by the vacuum pump 2 in the nucleic acid extract transfer mode is lower than the degree of decompression by the vacuum pump 2 in the sample discharge mode.

As a result, the nucleic acid extract 320 can be efficiently recovered with a simple structure.

Further, in the present embodiment, the vacuum pump 2 pressurizes the inside of the second container portion 12 to make the pressure in the second container portion 12 higher than the pressure in the first container portion 11, thereby forming the container 10. The pressure inside the second container unit 12 is adjusted in the nucleic acid extraction reagent holding mode in which the injected nucleic acid extraction reagent 310 is held on the capture unit 40.

As a result, the nucleic acid extraction reaction can be carried out in a state where the nucleic acid extraction reagent 310 is held on the capture unit 40 by adjusting the pressure in the second container unit 12, so that the nucleic acid extract 320 can be efficiently recovered. Can be done.

Further, in the present embodiment, the second injection port 22 is a through hole provided in the first container portion 11, and the through hole is provided with a rubber stopper 22a for closing the through hole. The 22a is made of a rubber material through which a needle for injecting the nucleic acid extraction reagent 310 into the first container portion 11 can penetrate.

Thereby, it is possible to reduce that the sample 210 and the nucleic acid extraction reagent 310 injected into the container 10 evaporate from the container 10 or leak from the container 10 at the time of stirring.

Further, in the present embodiment, the capture unit 40 has a main body portion 41 for capturing and holding the nucleic acid extraction target 220 contained in the sample 210, and a support portion 42 for supporting the main body portion 41. There is.

As a result, the nucleic acid extraction target 220 contained in the sample 210 can be captured in the main body 41.

In this case, at least a part of the support portion 42 may be made of metal. Alternatively, at least a part of the support portion 42 may be made of a high thermal conductive resin.

As a result, the heating / cooling unit 7 connected to the support portion 42 can efficiently heat or cool the sample 210, the nucleic acid extraction target 220, or the nucleic acid extraction reagent 310 on the main body 41.

Further, when the support portion 42 is made of a high thermal conductive resin and is not made of a metal, it is possible to reduce that the inspection is hindered by the metal itself or the corrosion of the metal.

Further, the surface of the support portion 42 of the capture portion 40 is preferably hydrophilic.

Thereby, the speed of capturing the nucleic acid extraction target 220 from the sample 210 can be improved. That is, the time required to capture the nucleic acid extraction target 220 from the sample 210 can be shortened.

Further, in the present embodiment, it is preferable that at least a part of the container 10 is made of a highly thermally conductive resin.

As a result, the heating / cooling unit 7 connected to the container 10 can efficiently heat or cool the sample 210, the nucleic acid extraction target 220, or the nucleic acid extraction reagent 310 on the main body 41. Further, by not using a metal as the container 10, it is possible to reduce that the inspection is hindered by the metal itself or the corrosion of the metal.

Further, in the present embodiment, the angle of the corner portion on the wall surface of the internal space of the first container portion 11 is preferably 90 degrees or more.

As a result, when the sample 210 and the nucleic acid extraction reagent 310 are injected into the first container portion 11, it is possible to prevent the sample 210 and the nucleic acid extraction reagent 310 from staying on the wall surface of the internal space of the first container portion 11. That is, the liquid loss of the sample 210 and the nucleic acid extraction reagent 310 can be reduced.

Further, in the present embodiment, the inner surface of the container 10 is preferably non-hydrophilic.

As a result, when the sample 210 and the nucleic acid extraction reagent 310 are injected into the first container portion 11, it is possible to prevent the sample 210 and the nucleic acid extraction reagent 310 from adhering to the wall surface of the internal space of the first container portion 11. .. That is, the liquid loss of the sample 210 and the nucleic acid extraction reagent 310 can be reduced.

Further, in the present embodiment, the nucleic acid extraction device 100 further includes a vibration device 3 for vibrating the container 10.

As a result, the nucleic acid extraction reagent 310 and the nucleic acid extraction target 220 can be agitated and mixed, so that the nucleic acid 230 can be efficiently extracted, and the sample 210 and the nucleic acid are placed on the wall surface of the internal space of the first container portion 11. It is possible to suppress the adhesion of the extraction reagent 310.

Further, in the nucleic acid extraction method in the present embodiment, the capture step S1 in which the capture unit 40 captures the nucleic acid extraction target 220 contained in the sample 210 by passing the sample 210 through the capture unit 40, and the nucleic acid in the capture unit 40. The nucleic acid extraction step S2 for extracting the nucleic acid 230 from the nucleic acid extraction target 220 captured by the capture unit 40 by introducing the extraction reagent 310, and the nucleic acid extract 320 containing the extracted nucleic acid 230 are passed through the capture unit 40 and collected. The recovery step S3 is included.

As a result, workability is simple, and the risk of impurities being mixed in the nucleic acid extract 320 can be significantly reduced. Therefore, the nucleic acid 230 can be extracted with high sensitivity and high accuracy.

(Modification example)
The nucleic acid extraction device 100, the nucleic acid extraction unit 1, the nucleic acid extraction method, and the like according to the present invention have been described above based on the embodiments, but the present invention is not limited to the above embodiments.

For example, in the above embodiment, the end of the pipe constituting the second discharge port 32 on the second container portion 12 side is flush with the inner wall of the second container portion 12, but the second discharge port 32 is used. At least a part of the constituent pipe on the second container portion 12 side may be projected inward of the second container portion 12 from the inner wall of the second container portion 12.

In this case, as shown in FIG. 7, a protruding portion 12a is provided so that the entire circumference of the end portion of the pipe constituting the second discharge port 32 on the second container portion 12 side protrudes from the inner wall of the second container portion 12. Alternatively, as shown in FIG. 8, the protruding portion so as to project only the upper portion of the end portion of the pipe constituting the second discharge port 32 on the second container portion 12 side from the inner wall of the second container portion 12. 12b may be provided.

In this way, by projecting at least a part of the pipe constituting the second discharge port 32 on the second container portion 12 side toward the inside of the second container portion 12 from the inner wall of the second container portion 12. When the nucleic acid extract 320 is moved from the first container 11 to the second container 12, it is possible to prevent the nucleic acid extract 320 from entering the second discharge port 32. That is, the protrusions 12a and 12b function as eaves, and the nucleic acid extract 320 can be prevented from entering the pipe of the second discharge port 32. Therefore, the nucleic acid extract 320 can be recovered with a small loss.

Further, in the nucleic acid extraction method in the above-described embodiment, the killed bacterium removing step may be performed to inactivate and remove the killed bacterium contained in the sample. The killed bacterium removal step is a step of reacting and inactivating the killed bacterium nucleic acid by introducing a killed bacterium removing reagent (for example, a reagent that inhibits the amplification of the killed bacterium nucleic acid) into the capture unit 40. This is performed after the capture step S1. Specifically, the killed bacterium removal step is performed between the capture step S1 and the nucleic acid extraction step S2.

In this case, after capturing the nucleic acid extraction target in the capture unit 40, the killed bacteria removing reagent is put into the container 10 from the first injection port 21 or the second injection port 22 to capture the nucleic acid extraction target microorganism. A dead bacterium removing reagent is introduced onto the part 40, and the mixture is allowed to stand for about 5 to 20 minutes while cooling. As a result, dead bacteria can be removed and only live bacteria can be collected in the capture unit 40. As the killed bacterium removing reagent, for example, "Viable Bacteria Selection Kit for PCR" manufactured by Takara Bio Inc. can be used. Further, in order to allow the killed bacteria removing reagent to permeate into the main body 41 (filter) of the trap 40, a step of lightly sucking by the vacuum pump 2 and a step of stirring may be added as appropriate. Further, if necessary, the killed bacteria removal step may be repeated several times. The killed bacteria removing reagent may be discharged from the second discharge port 32 or the like through the main body 41 after the killed bacteria removing step is completed. In this way, by performing the killed bacterium removal step after the capture step S1, only the nucleic acid of the live bacterium can be extracted.

Further, in the above embodiment, the container 10 is composed of a light-shielding member, but the present invention is not limited to this. For example, as shown in FIG. 9, at least a part of the container 10 may be made of a transparent material.

By making a part of the container 10 a transparent material in this way, it becomes possible to observe the inside of the container 10 and to irradiate the inside of the container 10 with light. In this case, for example, a light irradiation unit 8 for irradiating the inside of the container 10 with light can be provided in the reaction box 4 or in the vicinity of the reaction box 4. The light irradiation unit 8 is used, for example, in the above-mentioned killed bacteria removing step. Specifically, the reaction between the dead bacteria and the dead bacteria removing reagent can be promoted by introducing the killed bacteria removing reagent into the container 10 and allowing it to stand, and then irradiating with light by the light irradiation unit 8. As a result, dead bacteria can be efficiently removed.

Further, in the above embodiment, the first injection port 21 for injecting the sample into the container 10 and the second injection port 22 for injecting the nucleic acid extraction reagent into the container 10 are separately provided, but the present invention is limited to this. It's not a thing. Further, the first discharge port 31 for discharging the nucleic acid extract from the container 10 and the second discharge port 32 for discharging the sample from the container 10 are also provided separately, but the present invention is not limited to this.

For example, the first injection port 21 and the second injection port 22 may be configured as one injection port 23 as shown in FIG. That is, one injection port 23 that also serves as the first injection port 21 and the second injection port 22 may be provided in the container 10 (first container portion 11). A sample is injected or a nucleic acid extraction reagent is injected from the injection port 23. In addition to the sample 210 and the nucleic acid extraction reagent 310, other liquids such as a dead bacterium removing reagent may be injected from the injection port 23.

Further, the first discharge port 31 and the second discharge port 32 may also be configured as one discharge port 33 as shown in FIG. That is, one discharge port 33 that also serves as the first discharge port 31 and the second discharge port 32 may be provided in the container 10 (second container portion 12). The nucleic acid extract is discharged or the sample is discharged from the discharge port 33. In addition to the nucleic acid extract and the sample, other liquids introduced into the container 10 may be discharged from the discharge port 33.

In FIG. 10, both the injection port 23 and the discharge port 33 are set to one, but the present invention is not limited to this, and one of them may be divided into two as shown in FIG. 6A and the like.

Further, when each of the injection port 23 and the discharge port 33 is only one, as shown in FIG. 11, the sample is charged into the injection port 23 provided on the upstream side of the capture unit 40 via the valve 600. The cup 200 and the nucleic acid extraction reagent container 300 may be connected. Thereby, by controlling the valve 600, the flow of the sample from the sample charging cup 200 and the nucleic acid extraction reagent from the nucleic acid extraction reagent container 300 can be switched. Further, at the discharge port 33 provided on the downstream side of the capture unit 40, the waste liquid of the sample is collected by the pipe 5b and the collection container 500 for collecting the nucleic acid extract in the collection container 400 (not shown) via the valve 700. A pipe 2b for collecting (filter liquid) may be connected. Thereby, the flow of the nucleic acid extract and the waste liquid of the sample can be switched by controlling the valve 700.

Further, in the above embodiment, the sample 210 is directly passed through the capture unit 40, but the present invention is not limited to this. For example, the sample 210 may be passed through the pre-filter as a pretreatment before the sample 210 is passed through the capture unit 40. As an example, a pipe having a pre-filter may be inserted between the sample charging cup 200 and the first injection port 21. In this way, by passing the sample 210 through the pre-filter before passing it through the capture unit 40, it is possible to remove unnecessary components having a large size. This makes it possible to efficiently capture the nucleic acid extraction target 220 from the sample 210.

In addition, it is realized by arbitrarily combining the components and functions in the embodiment and the embodiment obtained by applying various modifications that can be thought of by those skilled in the art to each embodiment and modification, and within the range not deviating from the gist of the present invention. The form to be used is also included in the present invention.

1 Nucleic acid extraction unit 2 Vacuum pump (pressure regulator)
10 Container 11 1st container 12 2nd container 21 1st injection port 22 2nd injection port 23 Injection port (1st injection port, 2nd injection port)
22a Rubber stopper 31 1st discharge port 32 2nd discharge port 33 Discharge port (1st discharge port, 2nd discharge port)
40 Capture part 41 Main body part 42 Support part 100 Nucleic acid extractor 210 Specimen 220 Nucleic acid extraction target 230 Nucleic acid 310 Nucleic acid extraction reagent 320 Nucleic acid extract

Claims (12)

A nucleic acid extraction unit for extracting nucleic acid from a sample,
And a pressure adjusting unit,
The nucleic acid extraction unit
With the container
A first injection port for injecting a sample into the container,
A capture unit that captures at least a part of the nucleic acid extraction target contained in the sample injected from the first injection port, which is arranged in the container.
A second injection port for injecting a nucleic acid extraction reagent for extracting nucleic acid from the nucleic acid extraction target captured by the capture unit into the container, and
A first discharge port for discharging the nucleic acid extract containing the nucleic acid extracted from the nucleic acid extraction target by the nucleic acid extraction reagent from the container, and
It has a second discharge port for discharging the waste liquid of the sample from the container.
The container has a first container portion and a second container portion that divide the internal space of the container with the catching portion as a boundary.
The first injection port and the second injection port are provided in the first container portion.
The first discharge port and the second discharge port are provided in the second container portion.
The first injection port and the second injection port are provided separately or are configured as one injection port.
The first discharge port and the second discharge port are provided separately.
The capture unit has a filter having a plurality of pores smaller than the nucleic acid extraction target.
The second discharge port is provided at a position closer to the catching portion than the first discharge port, and also serves as a decompression port of the pressure adjusting portion.
The pressure adjusting section pressurizes the inside of the second container section to make the pressure inside the second container section higher than the pressure inside the first container section, thereby capturing the nucleic acid extraction reagent injected into the container. A nucleic acid extraction device that adjusts the pressure inside the second container in the nucleic acid extraction reagent holding mode held on the container. The first aspect of claim 1, wherein at least a part of the pipe constituting the second discharge port on the side of the second container portion projects inward of the second container portion from the inner wall of the second container portion. Nucleic acid extractor. The second injection port is a through hole provided in the first container portion, and is a through hole.
The through hole is provided with a rubber stopper that closes the through hole.
The nucleic acid extraction device according to claim 1 or 2 , wherein the rubber stopper is made of a rubber material through which a needle for injecting the nucleic acid extraction reagent into the first container portion can penetrate. The nucleic acid extraction device according to any one of claims 1 to 3 , wherein at least a part of the container is made of a transparent material. The nucleic acid extraction device according to any one of claims 1 to 4 , wherein the capture unit has a main body portion for capturing and holding the nucleic acid extraction target, and a support portion for supporting the main body portion. The nucleic acid extraction device according to claim 5 , wherein at least a part of the support portion is made of metal. The nucleic acid extraction device according to claim 5 , wherein at least a part of the support portion is made of a highly thermally conductive resin. The nucleic acid extraction device according to any one of claims 5 to 7 , wherein the surface of the support portion is hydrophilic. The nucleic acid extraction device according to any one of claims 1 to 8 , wherein at least a part of the container is made of a highly thermally conductive resin. The nucleic acid extractor according to any one of claims 1 to 9 , wherein the angle of the corner portion on the wall surface of the internal space of the first container portion is 90 degrees or more. The nucleic acid extraction device according to any one of claims 1 to 10 , wherein the inner surface of the container is non-hydrophilic. The nucleic acid extraction device according to any one of claims 1 to 11 , further comprising a vibrating device for vibrating the container.

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